Comparison of the metabolism of retinol delivered to human keratinocytes either bound to serum retinol-binding protein or added directly to the culture medium.
Retinol (vitamin A) circulates in the blood bound to retinol-binding protein (RBP), which is thought to be responsible for the delivery of the vitamin to target cells, including the basal cells of the skin (keratinocytes). The process by which keratinocytes acquire retinol from RBP remains controversial. A mechanism for retinol delivery to keratinocytes involving cell surface RBP receptors has been proposed, while other studies support an RBP receptor-independent process. To further explore retinol uptake we have used a model system of human foreskin keratinocytes cultured in serum-free media to compare the metabolism of [3H]retinol delivered to the cells either bound to RBP or added directly to the culture medium. The majority of the cell-associated radioactivity found in keratinocytes incubated for 0.5 to 24 h with either free or RBP bound [3H]retinol was present as [3H]retinyl ester irrespective of the mode of delivery. In keratinocytes incubated for 24 h with [3H]retinol added directly to the culture medium or bound to RBP, [3H]retinyl ester comprised 76 and 80%, respectively, of the total cell-associated radioactivity. Also, the relative cellular levels of the different retinyl esters species synthesized by the keratinocytes were the same whether the [3H]-retinol was delivered free or bound to RBP. Finally, the kinetics of loss (turnover) of cell-associated [3H]-retinol and [3H]retinyl esters from keratinocytes pre-labeled with [3H]retinol delivered free or bound to RBP was the same. Overall, this study demonstrates that the rate and extent of retinol esterification by keratinocytes and the types of esters synthesized are the same whether the vitamin is delivered to the cells free or bound to RBP and argues against RBP receptor-mediated delivery of retinol to specific sites on the plasma membrane that influence overall retinol metabolism.
Hodam
J R
JR
Department of Chemistry and Biochemistry, University of South Carolina, Columbia 29208, USA.
Creek
K E
KE
D002478:Cells, Cultured; D003470:Culture Media; D004337:Drug Carriers; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D012176:Retinoids / Q000302:isolation & purification / Q000378:metabolism; D012177:Retinol-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D054839:Retinol-Binding Proteins, Plasma; D012867:Skin / Q000166:cytology; D014801:Vitamin A / Q000378:metabolism*
D003470:Culture Media; D004337:Drug Carriers; D012176:Retinoids; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D014801:Vitamin A